DNA Transformation to PEG competent cells 1. Transfer 100µl of competent cells to a 2059 snap-cap tube on ice.
2. Add 10µl of ligation product or up to 0.5µgr of super-coiled plasmid.
3. Incubate 30’ at 4^oC (on ice)
4. Incubate 45 sec in 42 ^oC * do not exceed 45''
5. Incubate 2 min on ice
6. Add 0.9 ml of Room Temp. SOC medium to the tube
7. Incubate in shaker at 37^oC for 1 hour (up to 90min) * The purpose of this step is to let the bacteria recover from the heat shock prior to plating on agar plates. Thus no antibiotic is added to the medium during this step, and the medium should not be cold.
8. Seed on plates 100µl of bugs , then spin the bugs 30'', aspirate sup and leave about 100µl, re-suspend the bugs in the medium, and plate all. * make sure you add the right antibiotics to the agar plates - check vector AND bacterial strains specifications.